KR960705041A - ACTIVIN RECEPTORS-LIKE KINASE (ALK), BELONGING TO THE TGF RECEPTOR FAMILY AND / OR TO THE BMP RECEPTOR FAMILY - Google Patents

ACTIVIN RECEPTORS-LIKE KINASE (ALK), BELONGING TO THE TGF RECEPTOR FAMILY AND / OR TO THE BMP RECEPTOR FAMILY

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KR960705041A
KR960705041A KR1019960701386A KR19960701386A KR960705041A KR 960705041 A KR960705041 A KR 960705041A KR 1019960701386 A KR1019960701386 A KR 1019960701386A KR 19960701386 A KR19960701386 A KR 19960701386A KR 960705041 A KR960705041 A KR 960705041A
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엠. 우즈니 존
제이. 셀레스트 안토니
스코트 티즈 알.
노보루 야마지
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토마스 제이. 데스로져
제네틱스 인스티튜트 인코포레이티드
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Abstract

Novel serine/threonine receptor proteins and BMP receptor proteins are disclosed, as well as DNA molecules encoding said proteins and methods of using the receptor proteins. Further disclosed are truncated BMP receptor proteins and molecules which act as ligands to said BMP receptor proteins.

Description

TGF 수용체 족 및/또는 BMP 수용체 족에 속하는 액티빈 수용체 유사 키나제(ALK)(ACTIVIN RECEPTORS-LIKE KINASE (ALK), BELONGING TO THE TGF RECEPTOR FAMILY AND/OR TO THE BMP RECEPTOR FAMILY)ACTIVIN RECEPTORS-LIKE KINASE (ALK), BELONGING TO THE TGF RECEPTOR FAMILY AND / OR TO THE BMP RECEPTOR FAMILY

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

Claims (52)

BMP 수용체 단백질을 코드하는 DNA 서열을 함유한 단리된 DNA분자.An isolated DNA molecule containing a DNA sequence encoding a BMP receptor protein. 제1항에 있어서, DNA 서열이 하기로 구성된 군으로부터 선개되는 DNA 분자 : (a) SEQ IN NO : 1의 누클레오티드 61~1656; (b) SEQ IN NO; 3의 누클레오티드 247~1752; 및 (c) 긴박한 혼성화 조건하에 (a) 또는 (b) 중 어느 하나에 혼성화되며, 결합 분석에 있어서 BMP에 결합할 수 있는 BMP 수용체 단백질을 코드하는 서열.The DNA molecule of claim 1, wherein the DNA sequence is selected from the group consisting of: (a) nucleotides 61-1656 of SEQ IN NO: 1; (b) SEQ IN NO; Nucleotides 247-1752 of 3; And (c) a sequence that hybridizes to either (a) or (b) under strict hybridization conditions and that encodes a BMP receptor protein capable of binding to BMP in a binding assay. DNA 서열이 하기로 구성된 군으로부터 선택되는, 세린/트레오닌키나제 수용체 단백질을 코드하는 DNA 서열을 함유한 단리된 DNA 분자 : (a) SEQ IN NO : 5의 누클레오티드 474~2000; (b) SEQ IN NO : 7의 누클레오티드 80~1594; (c) SEQ IN NO : 9의 누클레오티드 85~1591; (d) 긴박한 혼성화 조건하에 (a)~(c)중 어느 하나에 혼성화되며, 세린/트레오닌 수용체 단백질을 코드하는 서열.An isolated DNA molecule containing a DNA sequence encoding a serine / threonine kinase receptor protein, wherein the DNA sequence is selected from the group consisting of: (a) nucleotides 474-2000 of SEQ IN NO: 5; (b) nucleotide 80-1594 of SEQ IN NO: 7; (c) nucleotides 85-1591 of SEQ IN NO: 9; (d) a sequence that hybridizes to any one of (a) to (c) under strict hybridization conditions and codes for a serine / threonine receptor protein. 제1항에 있어서, DNA 서열이 하기로 구성된 군으로부터 선택되는 DNA 분자 : (a) SEQ IN NO : 2의 아미노산 24~532를 코드하는 누클레오티드; (b) SEQ IN NO : 4의 아미노산 8~502를 코드하는 누클레오티드; (c) 긴박한 혼성화 조건하에 (a) 또는 (b)에 혼성화되며, 결합 분석에 있어서 BMP에 결합할 수 있는 BMP 수용체 단백질을 코드하는 서열.The DNA molecule of claim 1, wherein the DNA sequence is selected from the group consisting of: (a) a nucleotide encoding amino acids 24-532 of SEQ IN NO: 2; (b) a nucleotide encoding amino acids 8-502 of SEQ IN NO: 4; (c) a sequence that hybridizes to (a) or (b) under imminent hybridization conditions and encodes a BMP receptor protein capable of binding to BMP in a binding assay. DNA 서열이 하기로 구성된 군으로부터 선택되는, 세린/카나제 수용체 단백질을 코드하는 DNA 서열을 함유하는 단리된 DNA 분자 : (a) SEQ IN NO : 6의 아미노산 18~509를 코드하는 누클레오티드; (b) SEQ IN NO : 8의 아미노산 24~505를 코드하는 누클레오티드; (c) SEQ IN NO : 10의 아미노산 30~503를 코드하는 누클레오티드; (d) 긴박한 혼성화 조건하에 (a)~(c)중 어느 하나에 혼성화되며, 세린/트레오닌 카나제 수용체 단백질을 코드하는 서열.An isolated DNA molecule containing a DNA sequence encoding a serine / canase receptor protein, wherein the DNA sequence is selected from the group consisting of: (a) a nucleotide encoding amino acids 18-509 of SEQ IN NO: 6; (b) a nucleotide encoding amino acids 24-505 of SEQ IN NO: 8; (c) a nucleotide encoding amino acids 30-503 of SEQ IN NO: 10; (d) A sequence that hybridizes to any one of (a) to (c) under strict hybridization conditions and codes for a serine / threonine kinase receptor protein. 제1항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 1. 제2항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 2. 제4항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 4. 제5항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 5. 결합 분석에 있어서 BMP에 결합하는 능력을 특징으로 하는 끝이 잘린 BMP 수용체 단백질을 코드하는 서열을 가지며, 하기로 구성된 군으로부터 선택된 DNA 서열을 함유하는 단리된 DNA 분자 : (a) SEQ ID NO : 1의 누클레오티드 61~507; (b) SEQ ID NO : 2의 아미노산 1~149를 코드하는 누클레오티드; (c) SEQ IN NO :3의 누클레오티드 247~618; (d) SEQ ID NO : 4의 아미노산 8~124를 코드하는 누클레오티드; 및 (e) 자연적으로 발생하는 대립 유전자(allelic) 서열 및 (a), (b), (c) 또는 (d)에 대한 등가의 축퇴 코돈 서열.An isolated DNA molecule having a sequence encoding a truncated BMP receptor protein characterized by the ability to bind BMP in a binding assay and containing a DNA sequence selected from the group consisting of: (a) SEQ ID NO: 1 Nucleotides 61-507; (b) a nucleotide encoding amino acids 1-149 of SEQ ID NO: 2; (c) nucleotides 247-618 of SEQ IN NO: 3; (d) a nucleotide encoding amino acids 8-124 of SEQ ID NO: 4; And (e) naturally occurring allelic sequences and equivalent degenerate codon sequences for (a), (b), (c) or (d). 제10항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 10. 제10항의 DNA 분자에 대한 발현 조절 서열과 작동상 연관된 제10항의 DNA 분자를 함유하는 벡터.A vector containing a DNA molecule of claim 10 operatively associated with an expression control sequence for the DNA molecule of claim 10. 하기 단계로 이루어진 정제된 끝이 잘린 BMP 수용체 단백질의 제조방법 : (a) 끝이 잘린 BMP 수용체 단백질을 코드하는 누클레오티드 서열을 함유하는, 제10항에 따른 DNA 서열로 형질 전환된 숙주 세포를 배양배지중에 배양하고; 및 (b) 배양 배지로부터 끝이 잘린 BMP 수용체 단백질을 회수 및 정제한다.A method for producing a purified truncated BMP receptor protein comprising the following steps: (a) A culture medium for host cells transformed with the DNA sequence according to claim 10, which contains a nucleotide sequence encoding a truncated BMP receptor protein. Incubated in the air; And (b) recovering and purifying the truncated BMP receptor protein from the culture medium. SEQ IN NO : 2의 아미노산 24~149의 아미노산 서열을 함유하는 정제된 끝이 잘린 BMP 수용체 단백질.A purified truncated BMP receptor protein containing the amino acid sequence of amino acids 24-149 of SEQ IN NO: 2. SEQ IN NO : 4의 아미노산 8~124의 아미노산 서열을 함유하는 정제된 끝이 잘린 BMP 수용체 단백질.A purified truncated BMP receptor protein containing the amino acid sequences of amino acids 8-124 of SEQ IN NO: 4. 하기 단계에 의해 제조된 정제된 끝이 잘린 BMP 수용체 단백질 : (a) SEQ ID NO : 1의 누클레오티드 61~507의 누클레오티드 서열을 함유하는 DNA로 형질 전환된 세포를 배양 배지중에 배양하고; 및 (b) SEQ ID NO : 2로 구성된 군으로부터 선개된 서열중 아미노산 24~149의 아미노산 서열을 함유하는 단백질을 배양 배지로부터 회수 및 정제한다.Purified truncated BMP receptor protein prepared by the following steps: (a) culturing cells transformed with DNA containing the nucleotide sequences of nucleotides 61-507 of SEQ ID NO: 1 in culture medium; And (b) recovering and purifying from the culture medium a protein containing an amino acid sequence of amino acids 24 to 149 among the sequences opened from the group consisting of SEQ ID NO: 2. 하기 단계에 의해 제조된 정제된 끝이 잘린 BMP 수용체 단백질 : (a) SEQ ID NO : 3의 누클레오티드 247~618의 누클레오티드 서열을 함유하는 DNA로 형질 전환된 세포를 배양 배지중에 배양하고; 및 (b) SEQ ID NO : 4로 구성된 군으로부터 선택된 서열중 아미노산 8~124의 아미노산 서열을 함유하는 단백질을 배양배지로부터 회수 및 정제한다.Purified truncated BMP receptor protein prepared by the following steps: (a) culturing cells transformed with DNA containing the nucleotide sequences of nucleotides 247-618 of SEQ ID NO: 3 in culture medium; And (b) recovering and purifying the protein containing the amino acid sequence of amino acids 8-124 from the culture medium selected from the group consisting of SEQ ID NO: 4 from the culture medium. 하기 단계로 이루어진 끝이 달린 BMP 수용체 단백질의 제조 방법 :(a) BMP 수용체 단백질의 리간드 결합 부위를 코드하는 끝이 잘린 누클레오티드 서열을 함유하는, 끝이 잘린 BMP 수용체 단백질을 코드하는 DNA 서열로 형질 전환된 숙주 세포를 배양 배지중에 배양하고; 및 (b) 배양 배지로부터 BMP 수용체 단백질을 회수 및 정제한다.A method for producing an endless BMP receptor protein consisting of the following steps: (a) Transformation into a DNA sequence encoding a truncated BMP receptor protein containing a truncated nucleotide sequence encoding a ligand binding site of the BMP receptor protein Cultured host cells in culture medium; And (b) recover and purify the BMP receptor protein from the culture medium. 하기 단계로 이루어진 끝이 잘린 BMP 수용체 단백질의 제조 방법 : (a) BMP 수용체 단백질의 리간드결합 부위를 코드하는 끝이 잘린 누클레오티드 서열을 함유하는, 제11항에 따른 숙주 세포를 배양 배지중에 배양하고; 및 (b) 배양 배지로부터 BMP 수용체 단백질을 회수 및 정제한다.A method for preparing a truncated BMP receptor protein, comprising the steps of: (a) culturing a host cell according to claim 11 containing a truncated nucleotide sequence encoding a ligand binding site of a BMP receptor protein in culture medium; And (b) recover and purify the BMP receptor protein from the culture medium. 끝이 잘린 세린/트레오닌 키나제 수용체 단백질을 코드하는 서열을 가지며, 하기로 구성된 군으로부터 선택된 DNA 서열을 함유하는 단리된 DNA 분자 : (a) SEQ ID NO : 5의 누클레오티드 474~836; (b) SEQ ID NO : 6의 아미노산 1~121을 코드하는 누클레오티드; (c) SEQ ID NO : 7의 누클레오티드 80~445; (d) SEQ ID NO :8의 아미노산 1~122를 코드하는 누클레오티드; (e) SEQ ID NO :9의 누클레오티드 83~445; (f) SEQ ID NO : 10의 아미노산 1~121을 코드하는 누클레오티드; 및 (g) 자연적으로 발생하는 대립 유전자(allelic) 서열 및 (a)~(f)에 대한 등가의 축퇴 코돈 서열.An isolated DNA molecule having a sequence encoding a truncated serine / threonine kinase receptor protein and containing a DNA sequence selected from the group consisting of: (a) nucleotides 474-836 of SEQ ID NO: 5; (b) a nucleotide encoding amino acids 1-121 of SEQ ID NO: 6; (c) nucleotides 80-445 of SEQ ID NO: 7; (d) a nucleotide encoding amino acids 1-122 of SEQ ID NO: 8; (e) nucleotides 83-445 of SEQ ID NO: 9; (f) a nucleotide encoding amino acids 1-121 of SEQ ID NO: 10; And (g) naturally occurring allelic sequences and equivalent degenerate codon sequences for (a)-(f). 제20항의 DNA 분자로 형질 전환된 숙주 세포.A host cell transformed with the DNA molecule of claim 20. 제20항의 DNA 분자에 대한 발현 조절 서열과 작동상 연관된 제20항의 DNA 분자를 함유하는 벡터.A vector containing a DNA molecule of claim 20 operatively associated with an expression control sequence for the DNA molecule of claim 20. 하기 단계로 이루어진 정제된 끝이 잘린 세린/트레오닌 키나제 수용체 단백질의 제조 방법 :(a) 끝이 잘린 세린/트레오닌 키나제 수용체 단백질을 코드하는 누클레오티드 서열을 함유하는 제20항에 따른 DNA 서열로 형질 전환된 숙주세포를 배양 배지중에 배양하고; 및 (b) 배양 배지로부터 끝이 잘린 세린/트레오닌 키나제 수용체 단백질을 회수 및 정제한다.A method for preparing a purified truncated serine / threonine kinase receptor protein comprising the following steps: (a) transformed with a DNA sequence according to claim 20 containing a nucleotide sequence encoding a truncated serine / threonine kinase receptor protein Host cells are cultured in culture medium; And (b) recovering and purifying the truncated serine / threonine kinase receptor protein from the culture medium. SEQ ID NO : 6의 아미노산 1~121의 아미노산 서열을 함유하는 정제된 끝이 잘린 세린/트레오닌 키나제 수용체 단백질.A purified truncated serine / threonine kinase receptor protein comprising amino acid sequences of amino acids 1-121 of SEQ ID NO: 6. SEQ ID NO : 8의 아미노산 1~122의 아미노산 서열을 함유하는 정제된 끝이 잘린 세린/트레오닌 키나제 수용체 단백질.A purified truncated serine / threonine kinase receptor protein comprising the amino acid sequences of amino acids 1-122 of SEQ ID NO: 8. SEQ ID NO : 10의 아미노산 1~121의 아미노산 서열을 함유하는 정제된 끝이 잘린 세린/트레오닌 키나제 수용체 단백질.A purified truncated serine / threonine kinase receptor protein comprising the amino acid sequences of amino acids 1-121 of SEQ ID NO: 10. 하기 단계로 이루어진 끝이 잘린 세린/트레오닌 카나제 수용체 단백질의 제조방법 : (a) 세린/트레오닌 키나제 수용체 단백질의 리간드 결합 부위를 코드하는 끝이 누클레오티드 서열을 함유하는 끝이 잘린 세린/트레오닌 카나제수용체 단백질을 코드하는 DNA 서열로 형질 전환된 숙주 세포를 배양 배지중에 배양하고; 및 (b) 배양 배재로부터 끝이 잘린 세린/트레오닌 키나제 수용체 단백질을 회수 및 정제한다.A method for preparing a truncated serine / threonine kinase receptor protein comprising the following steps: Culturing the host cell transformed with the DNA sequence encoding the protein in the culture medium; And (b) recovering and purifying the truncated serine / threonine kinase receptor protein from the culture medium. 하기 단계로 이루어진 끝이 잘린 세린/트레오닌 키나제 수용체 단백질의 제조 방법 : (a) 세린/트레오닌 키나제 수용체 단백질의 리간드 결합 부위를 코드하는 끝이 잘린 누클레오티드 서열을 함유하는, 제21항에 따른 숙주 세포를 배양 배재중에 배양하고; 및 (b) 배양 배지로부터 끝이 잘린 세린/트레오닌 키나제 수용체 단백질을 회수 및 정제한다.A method for producing a truncated serine / threonine kinase receptor protein comprising the following steps: (a) A host cell according to claim 21 containing a truncated nucleotide sequence encoding a ligand binding site of a serine / threonine kinase receptor protein Culture in culture exclusion; And (b) recovering and purifying the truncated serine / threonine kinase receptor protein from the culture medium. 하기 단계로 이루어진 방법을 통해 단리된 BMP 수용체 단백질을 코드하는 DNA 분자 :(a) 탐침으로서 공지된 BMP 수용체 단백질의 리간드 결합 부위를 코드하는 DNA 단편을 제조하고; (b) 탐침으로서 (a)의 DNA 단편을 사용하여, 게놈 또는 cDNA 라이브러리를 스크린하고; (c) (b) 단계에서 확인된 탐침으로 혼성화 DNA 분자를 단리하고; 및 (d) (c) 단계의 DNA 분자로부터 BMP수용체 단백질을 코드하는 DNA 분자를 클론한다.A DNA molecule encoding a BMP receptor protein isolated by a method consisting of: (a) preparing a DNA fragment encoding a ligand binding site of a BMP receptor protein known as a probe; (b) screening the genome or cDNA library using the DNA fragment of (a) as a probe; (c) isolating hybridized DNA molecules with the probe identified in step (b); And (d) clones a DNA molecule encoding a BMP receptor protein from the DNA molecule of step (c). 공지된 BMP 수용체 단백질이 CFK1~23a 또는 CFK1~43a인 제29항의 BMP 수용체 단백질.The BMP receptor protein of claim 29, wherein the known BMP receptor protein is CFK1-23a or CFK1-43a. CFK1~23a의 DNA 서열을 함유하는 단리된 DNA 분자.An isolated DNA molecule containing the DNA sequence of CFK1-23a. CFK1~43a의 DNA 서열을 함유하는 단리된 DNA 분자.An isolated DNA molecule containing the DNA sequence of CFK1-43a. CFK1~10a의 DNA 서열을 함유하는 단리된 DNA 분자.An isolated DNA molecule containing the DNA sequence of CFK1-10a. W-101의 DNA 서열을 함유하는 단리된 DNA 분자.Isolated DNA molecule containing the DNA sequence of W-101. W-120의 DNA 서열을 함유하는 단리된 DNA 분자.Isolated DNA molecule containing the DNA sequence of W-120. 하나 이상의 BMP 수용체 단백질을 코드하는 DNA 서열을 함유하는 DNA 분자로 형질 전환된 세포를 포함하는 조성물.A composition comprising a cell transformed with a DNA molecule containing a DNA sequence encoding at least one BMP receptor protein. BMP 유사 모노클론성 항체, 수형 펩티드 BMP 유사체, 및 소형 유기 분자 BMP 유사체로 구성된 군으로부터 선택되는, 끝이 잘린 BMP 수용체 단백질에 결합되는 르간드.Ligand bound to a truncated BMP receptor protein selected from the group consisting of BMP-like monoclonal antibodies, male peptide BMP analogs, and small organic molecule BMP analogs. 제37항에 있어서, BMP 유사 모노클론성 항체를 함유하는 리간드.The ligand of claim 37 which contains a BMP-like monoclonal antibody. 제37항에 있어서, 소형 펩티드 BMP 유사체를 함유하는 리간드.The ligand of claim 37 containing a small peptide BMP analog. 제37항에 있어서, 소형 유기 분자 BMP 유사체를 함유하는 리간드.The ligand of claim 37 containing a small organic molecule BMP analog. 제20항의 끝이 잘린 BMP 수용체에 대한 리간드로서 사용하기 위해 1차로 확인된 화합물을 함유하는 약학적 조성물.A pharmaceutical composition containing a compound identified primarily for use as a ligand for a truncated BMP receptor of claim 20. 제41항에 있어서, BMP 유사 모노클론성 항체를 함유하는 조성물.42. The composition of claim 41, which contains a BMP like monoclonal antibody. 제41항에 있어서, 소형 펩티드 BMP 유사체를 함유하는 조성물.42. The composition of claim 41, wherein the composition contains small peptide BMP analogs. 제41항에 있어서, 소형 유기 분자 BMP 유사체를 함유하는 조성물.42. The composition of claim 41, wherein the composition contains small organic molecule BMP analogs. BMP 유사 모노클론성 항체, 소형 펩티드 BMP 유사체, 및 소형 유기 분자BMP 유사체로 구성된 군으로부터 선택되는, 제13항의 방법에 의해 제조된 끝이 잘린 BMP 수용체 단백질에 대한 리간드.A ligand for the truncated BMP receptor protein prepared by the method of claim 13, selected from the group consisting of BMP-like monoclonal antibodies, small peptide BMP analogs, and small organic molecule BMP analogs. 제45항에 있어서, BMP 유사 모노클론성 항체를 함유하는 리간드.46. The ligand of claim 45, wherein the ligand contains a BMP-like monoclonal antibody. 제45항에 있어서, 소형 펩티드 BMP 유사체를 함유하는 리간드.46. The ligand of claim 45, wherein the ligand contains a small peptide BMP analog. 제45항에 있어서, 소형 유기 분자 BMP 유사체를 함유하는 리간드.46. The ligand of claim 45, wherein the ligand contains small organic molecule BMP analogs. 제14항의 방법에 의해 제조된 끝이 잘린 BMP 수용체에 대한 리간드로서 사용하기 위해 1차로 확인된 화합물을 함유하는 약학적 조성물.A pharmaceutical composition containing a compound identified primarily for use as a ligand for a truncated BMP receptor prepared by the method of claim 14. 제49항에 있어서, BMP 유사 모노클론성 항체를 함유하는 조성물.The composition of claim 49 containing a BMP like monoclonal antibody. 제49항에 있어서, 소형 펩티드 BMP 유사체를 함유하는 조성물.The composition of claim 49 containing a small peptide BMP analog. 제49항에 있어서, 소형 유기 분자 BMP 유사체를 함유하는 조성물.The composition of claim 49 containing small organic molecule BMP analogs. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
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